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British Journal of Cancer logoLink to British Journal of Cancer
. 1999 Dec 8;82(1):171–177. doi: 10.1054/bjoc.1999.0896

Application of an original RT-PCR–ELISA multiplex assay for MDR1 and MRP, along with p53 determination in node-positive breast cancer patients

J M Ferrero 1, M C Etienne 1, J L Formento 1, M Francoual 1, P Rostagno 1, I Peyrottes 1, F Ettore 1, E Teissier 1, P Leblanc-Talent 1, M Namer 1, G Milano 1
PMCID: PMC2363171  PMID: 10638986

Abstract

The long-term prognostic value of tumoural MDR1 and MRP, along with p53 and other classical parameters, was analysed on 85 node-positive breast cancer patients receiving anthracycline-based adjuvant therapy. All patients underwent tumour resection plus irradiation and adjuvant chemotherapy (the majority receiving fluorouracil–epirubicin–cyclophosphamide). Median follow-up for the 54 alive patients was 7.8 years. Mean age was 53.7 years (range 28–79) and 54 patients were post-menopausal. MDR1 and MRP expression were quantified according to an original reverse transcription polymerase chain reaction multiplex assay with colourimetric enzyme-linked immunosorbent assay detection(β2-microglobulin as control). P53 protein was analysed using an immunoluminometric assay (Sangtec). MDR1 expression varied within an 11-fold range (mean 94, median 83), MRP within a 45-fold range (mean 315, median 242) and p53 protein from the limit of detection (0.002 ng mg−1) up to 35.71 ng mg−1(mean 1.18, median 0.13 ng mg−1). P53 protein was significantly higher in oestrogen receptor (ER)-negative than in ER-positive tumours (P = 0.039). The higher the p53, the lower the MDR1 expression (P = 0.015, r = –0.27). P53 was not linked to progesterone receptor (PR) status, S phase fraction, or MRP. Significantly greater MDR1 expression was observed in grade I tumours (P = 0.029). No relationship was observed between MDR1 and MRP. Neither MDR1 nor MRP was linked to ER or PR status. Unlike MDR1, MRP was correlated with the S phase: the greater the MRP, the lower the S phase (P = 0.006, r = –0.42). Univariate Cox analyses revealed that MDR1, MRP, p53 and S phase had no significant influence on progression-free or specific survival. A tendency suggested that the greater the p53, the shorter the progression-free survival (P = 0.076 as continuous and 0.069 as dichotomous). © 2000 Cancer Research Campaign

Keywords: p53, MDR1, MRP, breast cancer, multiplex assay

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Selected References

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  1. Abbaszadegan M. R., Futscher B. W., Klimecki W. T., List A., Dalton W. S. Analysis of multidrug resistance-associated protein (MRP) messenger RNA in normal and malignant hematopoietic cells. Cancer Res. 1994 Sep 1;54(17):4676–4679. [PubMed] [Google Scholar]
  2. Allred D. C., Clark G. M., Elledge R., Fuqua S. A., Brown R. W., Chamness G. C., Osborne C. K., McGuire W. L. Association of p53 protein expression with tumor cell proliferation rate and clinical outcome in node-negative breast cancer. J Natl Cancer Inst. 1993 Feb 3;85(3):200–206. doi: 10.1093/jnci/85.3.200. [DOI] [PubMed] [Google Scholar]
  3. BLOOM H. J., RICHARDSON W. W. Histological grading and prognosis in breast cancer; a study of 1409 cases of which 359 have been followed for 15 years. Br J Cancer. 1957 Sep;11(3):359–377. doi: 10.1038/bjc.1957.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barrand M. A., Heppell-Parton A. C., Wright K. A., Rabbitts P. H., Twentyman P. R. A 190-kilodalton protein overexpressed in non-P-glycoprotein-containing multidrug-resistant cells and its relationship to the MRP gene. J Natl Cancer Inst. 1994 Jan 19;86(2):110–117. doi: 10.1093/jnci/86.2.110. [DOI] [PubMed] [Google Scholar]
  5. Beck J., Bohnet B., Brügger D., Bader P., Dietl J., Scheper R. J., Kandolf R., Liu C., Niethammer D., Gekeler V. Multiple gene expression analysis reveals distinct differences between G2 and G3 stage breast cancers, and correlations of PKC eta with MDR1, MRP and LRP gene expression. Br J Cancer. 1998;77(1):87–91. doi: 10.1038/bjc.1998.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beck W. T., Grogan T. M., Willman C. L., Cordon-Cardo C., Parham D. M., Kuttesch J. F., Andreeff M., Bates S. E., Berard C. W., Boyett J. M. Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors: consensus recommendations. Cancer Res. 1996 Jul 1;56(13):3010–3020. [PubMed] [Google Scholar]
  7. Castillo L., Milano G., Santini J., Demard F., Pierrefite V. Analysis of retinoic acid receptor beta expression in normal and malignant laryngeal mucosa by a sensitive and routine applicable reverse transcription-polymerase chain reaction enzyme-linked immunosorbent assay method. Clin Cancer Res. 1997 Nov;3(11):2137–2142. [PubMed] [Google Scholar]
  8. Chin K. V., Ueda K., Pastan I., Gottesman M. M. Modulation of activity of the promoter of the human MDR1 gene by Ras and p53. Science. 1992 Jan 24;255(5043):459–462. doi: 10.1126/science.1346476. [DOI] [PubMed] [Google Scholar]
  9. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  10. Dexter D. W., Reddy R. K., Geles K. G., Bansal S., Myint M. A., Rogakto A., Leighton J. C., Goldstein L. J. Quantitative reverse transcriptase-polymerase chain reaction measured expression of MDR1 and MRP in primary breast carcinoma. Clin Cancer Res. 1998 Jun;4(6):1533–1542. [PubMed] [Google Scholar]
  11. Filipits M., Suchomel R. W., Dekan G., Haider K., Valdimarsson G., Depisch D., Pirker R. MRP and MDR1 gene expression in primary breast carcinomas. Clin Cancer Res. 1996 Jul;2(7):1231–1237. [PubMed] [Google Scholar]
  12. Gasparini G., Pozza F., Harris A. L. Evaluating the potential usefulness of new prognostic and predictive indicators in node-negative breast cancer patients. J Natl Cancer Inst. 1993 Aug 4;85(15):1206–1219. doi: 10.1093/jnci/85.15.1206. [DOI] [PubMed] [Google Scholar]
  13. Iacopetta B., Grieu F., Powell B., Soong R., McCaul K., Seshadri R. Analysis of p53 gene mutation by polymerase chain reaction-single strand conformation polymorphism provides independent prognostic information in node-negative breast cancer. Clin Cancer Res. 1998 Jul;4(7):1597–1602. [PubMed] [Google Scholar]
  14. Lautier D., Canitrot Y., Deeley R. G., Cole S. P. Multidrug resistance mediated by the multidrug resistance protein (MRP) gene. Biochem Pharmacol. 1996 Oct 11;52(7):967–977. doi: 10.1016/0006-2952(96)00450-9. [DOI] [PubMed] [Google Scholar]
  15. Levesque M. A., Yu H., Clark G. M., Diamandis E. P. Enzyme-linked immunoabsorbent assay-detected p53 protein accumulation: a prognostic factor in a large breast cancer cohort. J Clin Oncol. 1998 Aug;16(8):2641–2650. doi: 10.1200/JCO.1998.16.8.2641. [DOI] [PubMed] [Google Scholar]
  16. Linn S. C., Giaccone G., van Diest P. J., Blokhuis W. M., van der Valk P., van Kalken C. K., Kuiper C. M., Pinedo H. M., Baak J. P. Prognostic relevance of P-glycoprotein expression in breast cancer. Ann Oncol. 1995 Sep;6(7):679–685. doi: 10.1093/oxfordjournals.annonc.a059284. [DOI] [PubMed] [Google Scholar]
  17. Lowe S. W., Bodis S., McClatchey A., Remington L., Ruley H. E., Fisher D. E., Housman D. E., Jacks T. p53 status and the efficacy of cancer therapy in vivo. Science. 1994 Nov 4;266(5186):807–810. doi: 10.1126/science.7973635. [DOI] [PubMed] [Google Scholar]
  18. Lowe S. W., Ruley H. E., Jacks T., Housman D. E. p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell. 1993 Sep 24;74(6):957–967. doi: 10.1016/0092-8674(93)90719-7. [DOI] [PubMed] [Google Scholar]
  19. McDonagh T. A., Robb S. D., Murdoch D. R., Morton J. J., Ford I., Morrison C. E., Tunstall-Pedoe H., McMurray J. J., Dargie H. J. Biochemical detection of left-ventricular systolic dysfunction. Lancet. 1998 Jan 3;351(9095):9–13. doi: 10.1016/s0140-6736(97)03034-1. [DOI] [PubMed] [Google Scholar]
  20. Muss H. B., Thor A. D., Berry D. A., Kute T., Liu E. T., Koerner F., Cirrincione C. T., Budman D. R., Wood W. C., Barcos M. c-erbB-2 expression and response to adjuvant therapy in women with node-positive early breast cancer. N Engl J Med. 1994 May 5;330(18):1260–1266. doi: 10.1056/NEJM199405053301802. [DOI] [PubMed] [Google Scholar]
  21. Namer M., Ramaioli A., Fontana X., Etienne M. C., Héry M., Jourlait A., Milano G., Frenay M., François E., Lapalus F. Prognostic value of total cathepsin D in breast tumors. A possible role in selection of chemoresistant patients. Breast Cancer Res Treat. 1991 Oct;19(2):85–93. doi: 10.1007/BF01980938. [DOI] [PubMed] [Google Scholar]
  22. Noonan K. E., Beck C., Holzmayer T. A., Chin J. E., Wunder J. S., Andrulis I. L., Gazdar A. F., Willman C. L., Griffith B., Von Hoff D. D. Quantitative analysis of MDR1 (multidrug resistance) gene expression in human tumors by polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7160–7164. doi: 10.1073/pnas.87.18.7160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nooter K., Brutel de la Riviere G., Look M. P., van Wingerden K. E., Henzen-Logmans S. C., Scheper R. J., Flens M. J., Klijn J. G., Stoter G., Foekens J. A. The prognostic significance of expression of the multidrug resistance-associated protein (MRP) in primary breast cancer. Br J Cancer. 1997;76(4):486–493. doi: 10.1038/bjc.1997.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pastan I., Gottesman M. Multiple-drug resistance in human cancer. N Engl J Med. 1987 May 28;316(22):1388–1393. doi: 10.1056/NEJM198705283162207. [DOI] [PubMed] [Google Scholar]
  25. Raybaud-Diogène H., Tétu B., Morency R., Fortin A., Monteil R. A. p53 overexpression in head and neck squamous cell carcinoma: review of the literature. Eur J Cancer B Oral Oncol. 1996 May;32B(3):143–149. doi: 10.1016/0964-1955(95)00095-x. [DOI] [PubMed] [Google Scholar]
  26. Remvikos Y., Beuzeboc P., Zajdela A., Voillemot N., Magdelénat H., Pouillart P. Correlation of pretreatment proliferative activity of breast cancer with the response to cytotoxic chemotherapy. J Natl Cancer Inst. 1989 Sep 20;81(18):1383–1387. doi: 10.1093/jnci/81.18.1383. [DOI] [PubMed] [Google Scholar]
  27. Romain S., Formento J. L., Guirou O., Francoual M., Milano G., Martin P. M. Determination of oestrogen receptors by enzyme immunoassay. Technical differences between laboratories and their consequences. Eur J Cancer. 1994;30A(6):740–746. doi: 10.1016/0959-8049(94)90285-2. [DOI] [PubMed] [Google Scholar]
  28. Shimamura A., Fisher D. E. p53 in life and death. Clin Cancer Res. 1996 Mar;2(3):435–440. [PubMed] [Google Scholar]
  29. Silvestrini R., Benini E., Veneroni S., Daidone M. G., Tomasic G., Squicciarini P., Salvadori B. p53 and bcl-2 expression correlates with clinical outcome in a series of node-positive breast cancer patients. J Clin Oncol. 1996 May;14(5):1604–1610. doi: 10.1200/JCO.1996.14.5.1604. [DOI] [PubMed] [Google Scholar]

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